Expression of miR-146b in peripheral blood serum and aortic tissues in patients with acute type Stanford A aortic dissection and its clinical significance / 中南大学学报(医学版)

Objective:To explore expression of miR-146b in peripheral blood serum and aortic wall tissues in patients with acute Stanford type A aortic dissection (TAAD),and to discuss the significance and underlying mechanisms.Methods:The subjects were divided into a control group (excluded relative aortic diseases) (n=23) and a TAAD group (n=27).The miR-146b levels of serum and aortic wall tissues were detected by quantitative real-time PCR (qRT-PCR).Serum miR-146b and aortic wall tissues miR-146b were compared among different risk TAAD groups.The correlations between miR-146b and severity of aortic dissection were analyzed.MiR-146b related target genes were predicted by the DIANA LAB-TarBase 6.0 and TargetScan.Results:The expression levels of miR-146b in the serum and aortic wall tissues in the TAAD group were significantly elevated compared with those in the control group (P＜0.001).Compared with the mild risk group,the miR-146b levels of serum and aortic wall tissues were significantly higher in the moderate risk and severe risk groups (P＜0.05).The expression of miR-146b was positively correlated with the risk severity of TAAD patients (r=0.862,0.872;P＜0.05).Nuclear factor kappa B1 (NF-κB1),tumor necrosis factor receptor-associated factor 6 (TRAF6),matrix metalloproteinase 16 (MMP16) and actin alpha 2 (ACTA2) were miR-146b related target genes.Conclusion:The upregulation of miR-146b in peripheral blood serum and aortic wall tissues may contribute to the pathogenesis of TAAD and the severity of this disease.

Expression and function of miRNA211 in human cutaneous melanoma / 中华皮肤科杂志

Objective To determine the expression of miRNA211 (miR-211) in the development of malignant melanoma,and to investigate the correlation between miR-211 and its target molecule,matrix metalloproteinase 16 (MMP-16).Methods Cultured A375 melanoma cells were divided into 3 groups:miR-211 overexpression group and mock-vehicle group transfected with miR-211 mimics and empty vehicle respectively,and negative control group receiving no treatment.TaqMan fluorescence-based quantitative PCR was performed to determine the expression of miR-211 in HER1 primary melanocytes,A375,C32 and G361malignant melanoma cell lines,as well as in nevus tissues (n =18) and melanoma tissues (n =41),and to evaluate changes of MMP-16 mRNA expression in A375 cells before and after the overexpression of miR-211.Sulforhodamine B (SRB) assay and flow cytometry were conducted to evaluate cellular proliferative activity and determine cell cycle distribution respectively,and methylcellulose assay and Transwell assay to evaluate colony formation and cell migration abilities respectively.The size of selected colonies was used to represent colony formation ability,while the ratio of the number of migrating cells to that of non-migrating cells to represent cell migration ability.Results There were significant differences in the expression level of miR-211 among the G361,C32 and A375 cells (0.09 ± 0.02 vs.0.000 52 ± 0.000 20 vs.0.000 03 ± 0.000 01,F =10 410,P ＜ 0.01).The expression of miR-21 1 was significantly decreased in melanoma tissues compared with nevus tissues (0.17 ± 0.03 vs.0.87 ± 0.08,t =9.118,P ＜ 0.01).No significant differences were observed in cellular proliferative activity or cell cycle distribution among the miR-211 overexpression group,mock-vehicle group and negative control group.Compared with the mock-vehicle group,the miR-211 overexpression group showed significantly suppressed colony formation (0.49 ± 0.05 vs.0.85 ± 0.09,t =2.19,P ＜ 0.05) and cell migration (0.49 ± 0.06 vs.0.82 ± 0.09,t =3.15,P ＜ 0.05) abilities,while no significant difference was observed between the mock-vehicle group and negative control group.Additionally,the mRNA expression of MMP-16 significantly decreased in the miR-211 overexpression group compared with the mock-vehicle group after transfeetion (24 hours:0.33 ± 0.02 vs.0.91 ± 0.03,t =11.30,P ＜ 0.01;48 hours:0.52 ± 0.01 vs.0.96 ± 0.02,t =5.02,P ＜ 0.05;72 hours:0.71 ± 0.01 vs.0.97 ± 0.03,t =3.85,P ＜ 0.05),with no significant difference between the mock-vehicle group and negative control group at the above time points.Conclusions miR-211 was lowly expressed in both malignant melanoma cells and tissues,and it could inhibit both anchorage-independent growth and migration of melanoma cells.After up-regulation of miR-211 expression,the mRNA expression of MMP-16 decreased in A375 cells,suggesting that MMP-16 may be a downstream target of miR-211,and can influence melanoma metastasis.

miR-26a suppresses gastric cancer cell invasion by targeting MMP16 / 中国癌症杂志

Background and purpose:Invasion and metastasis lead to poor prognosis in gastric cancer. In this study, we investigated the potential function of miR-26a in gastric cancer.Methods:Real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of miR-26a in gastric cancer cells.In vitro CCK-8 assay, cloning formation assay and Matrigel-Transwell assay were used to evaluate the proliferation, migration and invasion of gastric cancer cells. A luciferase reporter assay was also conducted to confirm that matrix metallo-proteinase-16 (MMP16) is a direct target of miR-26a.Results:miR-26a was down-regulated in gastric cancer tissues compared with that in non-cancerous tissues. Functional studies showed that miR-26a inhibited cell proliferation, col-ony formation, cell motility and invasion. However, miR-26a had no effect on cell proliferation. We also characterized MMP16 as a direct target of miR-26a. We showed that knocking down MMP16 in gastric cancer cells signiifcantly de-creased MMP16 expression and inhibited cell invasion, whereas ectopic MMP16 expression signiifcantly abrogated the suppressed cell invasion induced by miR-26a.Conclusion:miR-26a suppresses gastric cancer cell invasion by targeting MMP16. miR-26a could represent a potential therapeutic target for gastric cancer.

miR-216a-5p inhibits invasion ability in human lung cancer cells by down-regulation of MMP16 expression / 中国癌症杂志

Background and purpose:MicroRNA (miRNA) belongs to a class of 19 to 30 nucleotide-long, endogenous noncoding RNA expressed in eukaryotes and predominantly inhibits gene expression at the post-transcriptional level. The miRNAs play critical roles in cell proliferation and differentiation, apoptosis, metabolism, and immune regulation. This study aimed to detect the expression of miR-216a-5p in lung cancer tissues and lung cancer cell lines, and to discuss the effects of miR-216a-5p on the invasion ability of lung cancer cells and the mechanism.Methods:Quantitative real-time PCR (qRT-PCR) was used to detect the expression of miR-216a-5p in lung cancer tissues of 55 cases and 7 lung cancer cell lines. Three lung cancer cell lines of A549, 95D and H460 were transiently transfected by miR-216a-5p, and Transwell was used to detect the effects of miR-216a-5p on the invasion of lung cancer cell lines. The dual luciferase reporter plasmids containing the miR-216a-5p candidate target gene and the gene of matrix metalloproteinase 16 (MMP16) were predicted and constructed. qRT-PCR and Western blot were used to detect the changes in mRNA and protein levels of target geneMMP16 by miR-216a-5p. The interference of MMP16 by siRNA and up-regulation miR-216a-5p by transfection were compared on the invasion of lung cancer cells.Results:The miR-216a-5p expression levels were all signiifcantly reduced in 90.91% (50 of 55 patients) tumor tissues compared with corresponding adjacent normal lung tissues (P<0.05). The miR-216a-5p expression levels were only 7.00%-32.00%in 7 lung cancer cells compared with the control group (P<0.05). Up-regulation of the expression of miR-216a-5p inhibited the invasion of lung cancer cells; interference of MMP16 by siRNA, as well as up-regulating miR-216a-5p by transfection, inhibited the expression of MMP16 in lung cancer leading to inhibition of the invasion of lung cancer cells. Conclusion:miR-216a-5p can be a candidate marker in clinical diagnosis and it can inhibit the invasion of lung cancer cells by down-regulating the expression of MMP16.

Expression and biological role of Matrix metalloproteinases 16 in esophagus squamous cell carcinoma / 中国癌症杂志

Background and purpose:Esophageal carcinoma is one of main malignancies with rapid course and a poor prognosis in China. The reasons of poor overall survival are the invasion and metastasis of the tumor. Matrix metalloproteinase (MMPs) play essential roles in promoting tumor invasion and metastasis. In this study, we aimed to investigate the expression and functional signiifcance of matrix metalloproteinase 16(MMP-16) in esophageal squamous cell carcinoma (ESCC). We expect to ifnd a lead molecule for the beneift of early detecting tumor and the development of novel treatment of ESCC. Methods:The expression levels of MMP-16 protein and mRNA in human ESCC and the matched normal tissues were determined by immunohistochemistry, Western blot and Real-Time PCR (RT-PCR). The stable Ec109 cell line with MMP-16 knockdown and negative controls were established by RNA interference technology. The cell migration, invasion, proliferation and cell apoptosis of MMP-16 in stable interfered Ec109 cell line was examined by cell counting, scratch test, Transwell test and lfow cytometry assays. The data were analyzed by t test. Results:MMP-16 protein was downregulated in cancerous group compared with the matched normal tissue and correlated with the clinical features of histological differentiation (P0.05). Conclusion: MMP-16 is downregulated in human ESCC tissues. The cell migration and invasion is promoted by interference of MMP-16 in Ec109, while the cell apoptosis is inhibited. MMP-16 may be considered as a target gene for therapy of ESCC.